With recent technical advances, multiple important signaling pathways that may be the causes of human malignancy have continuously been discovered and dissected. The vast majority of these signaling pathways involve reversible protein phosphorylation, and the information on the location and dynamics of phosphorylation provides important mechanisms on how the signaling networks function and interact. While translational research gradually shifts from lab models to clinical samples, with the ultimate goal of identifying cancer biomarkers, a simple and reliable phosphorylation assay method is still missing for routine detection of phosphorylation in complex and typically heterogeneous clinical samples. Through this NIH SBIR Phase I study we will develop soluble nanopolymer-based reagents, termed PolyMAC (Polymer-based Metal Ion Affinity Capture), into commercial products for the highly efficient isolation of phosphopeptides. This novel design takes advantage of not only the properties of multifunctionalized nanoparticles, but more importantly, the soluble nature of the molecule, allowing for the chelation of a limited amount of phosphopeptides in the solution phase for optimum efficiency and maximum yield. We propose to finalize optimization and scalability of the reagents as commercial products. In addition, high throughput formats for comprehensive phosphoproteomic analyses will be developed to address needs of many cancer biology and proteomics research labs/facilities. PUBLIC HEALTH RELEVANCE: Protein phosphorylation relates to the onset and development of many cancer types and a highly efficient technology for phosphorylation analysis is critical for cancer research. This NIH SBIR will support an effort to develop an innovative technology into commercial products that equip researchers with powerful tools and new directions to combat the devastating disease.